GENE TRANSFER IN LEUKOCYTE ADHERENCE DEFICIENCY

白细胞粘附缺陷中的基因转移

• 批准号: 2148762
• 负责人: DENNIS DURAND HICKSTEIN
• 金额: $ 11.04万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-09-15 至 1998-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2148762
• 关键词: CD antigens; Retroviridae; bone marrow; bone marrow transplantation; gene expression; gene targeting; gene therapy; genetic transduction; hematopoietic stem cells; human subject; integrins; laboratory mouse; leukocyte adhesion molecules; leukocyte disorder; protein structure; tissue /cell culture; transfection /expression vector

The aim of this proposal is to use retroviral vectors to direct the expression of the leukocyte integrin CD18 subunit in vitro in peripheral blood progenitor and bone marrow cells from normal individuals and from children with leukocyte adherence deficiency or LAD, and in vivo in bone marrow cells from mice in which the CD18 has been disrupted by gene targeting. Children with LAD experience recurrent, life-threatening bacterial infections due to the inability of the leukocytes from these children to adhere to the vessel wall and migrate to the site of infection. The clinical manifestations of the disease stem from the inability of leukocytes from children with LAD to express the members of the leukocyte integrin CD11/CD18 molecules on the cell surface due to defects in the CD18 subunit. Leukocyte adherence deficiency is a candidate disease for human gene therapy based upon gene transfer into hematopoietic progenitor cells and stem cells in that: l) the disease is life-threatening in the severe deficiency form, 2) except for bone marrow transplantation there is no treatment other than supportive therapy, 3) transduction of a normal CD18 gene into LAD EBV B-cells has been shown to correct the biochemical and functional defect, and 4) based upon gene expression by leukocytes in the moderate deficiency phenotype of LAD, 5% of normal levels of CD11/CD18 expression appears to be sufficient to correct the severe clinical manifestations of the disease. The current studies are designed to determine the optimal conditions for using retroviral vectors to restore CD18 expression in peripheral blood progenitors and bone marrow cells from LAD children with the ultimate goal of long-term disease correction by CD18 gene transfer into hematopoietic stem cells. In these studies we aim to: 1) determine the conditions required for retroviral vectors LgCD18SN and LgCD18, encoding the human CD18 subunit, to produce high efficiency gene transduction and gene expression of the CD18 subunit in peripheral blood progenitor cells and bone marrow cells from normal individuals and from children with LAD; 2) to determine the efficacy, efficiency, and safety of transduction of the human CD18 subunit into bone marrow cells from mice in whom the CD18 subunit has been disrupted by gene targeting. LAD is an attractive model for human gene therapy in that transduction of peripheral blood progenitor cells and bone marrow cells with CD18-encoding vectors ex vivo followed by re-infusion of the transduced cells may be major therapeutic benefit to children with LAD. During acute episodes of infection the granulocytes and monocytes which differentiate from the CD18-transduced progenitor cells may traffic to the inflammatory site and participate in the host response to the infectious agent. Thus, transduction of a hematopoietic stem cell may not be required for a clinical response in LAD.

该建议的目的是使用逆转录病毒向量指导 白细胞整合素CD18亚基在外周的体外表达 来自正常个体的血祖和骨髓细胞以及 白细胞依从性缺乏或小伙子的孩子，骨骼中的体内 CD18被基因破坏的小鼠的骨髓细胞 定位。有小伙子的孩子经历复发，威胁生命 细菌感染由于这些白细胞无法从中引起的 儿童遵守船只墙并迁移到 感染。该疾病的临床表现源于 小伙子儿童无法表达白细胞的成员 由于细胞表面上的白细胞整合素CD11/CD18分子由于 CD18亚基中的缺陷。 白细胞依从性缺乏是人类基因的候选疾病 基于基因转移到造血祖细胞和 干细胞在其中：l）这种疾病在严重的 缺陷形式，2）除骨髓移植外 除支持治疗以外的治疗，3）正常CD18的转导 已显示进入LAD EBV B细胞的基因可以纠正生化和 功能缺陷和4）基于白细胞的基因表达 LAD的中等缺陷表型，CD11/CD18的正常水平的5％ 表达似乎足以纠正严重的临床 疾病的表现。 目前的研究旨在确定 使用逆转录病毒载体在外周血中恢复CD18的表达 来自LAD儿童的祖细胞和骨髓细胞具有最终目标 通过CD18基因转移到造血的长期疾病校正 干细胞。在这些研究中，我们的目的是：1）确定条件 逆转录病毒载体LGCD18SN和LGCD18所需 CD18亚基，产生高效率基因转导和基因 CD18亚基在外周血祖细胞和 来自正常个体和LAD儿童的骨髓细胞； 2） 确定转导的功效，效率和安全性 人CD18亚基进入CD18的小鼠的骨髓细胞 亚基受到基因靶向的破坏。 LAD是人类基因疗法的有吸引力的模型 用CD18编码的外周血祖细胞和骨髓细胞 载体在体内，然后再输入转导的细胞可能是 对LAD儿童的主要治疗益处。在急性情节中 感染与区分的粒细胞和单核细胞 CD18转导的祖细胞可能会访问炎症部位，并 参与对传染剂的宿主反应。因此， 可能不需要造血干细胞的转导 LAD中的临床反应。